Preparation and characterization of an 125I-labeled Sendai virus ligand
| Autor: | Lucy Credle, Hugh C. McDonald, Albert August Luderer, Donna M. Hess |
|---|---|
| Rok vydání: | 1980 |
| Předmět: |
Erythrocytes
Hemagglutination viruses Biophysics Radioimmunoassay Biochemistry Virus Cell Line Sepharose Iodine Radioisotopes Radioligand Assay Murine leukemia virus Animals Humans Horses Molecular Biology Sheep biology Chemistry Cell Biology biology.organism_classification Ligand (biochemistry) Fetuin Molecular biology Sendai virus Lymphoproliferative Disorders Parainfluenza Virus 1 Human Isotope Labeling biology.protein Receptors Virus alpha-Fetoproteins Neuraminidase |
| Zdroj: | Analytical biochemistry. 106(1) |
| ISSN: | 0003-2697 |
| Popis: | In an attempt to prepare a radioviral ligand that was effective both as an antigen in binding to antibody and as a ligand that effectively binds to receptor-bearing cells, inactivated Sendai virus was adsorbed to immobilized fetuin at 4°C and recovered by temperature elevation at 37°C. The eluted virus was iodinated using the chloramine-T procedure and free iodine and labeled virus were separated on a Sepharose 4B column. Radiolabeted virus contained 1000 to 5000 cpm per hemagglutination unit and over 90% of the counts were sedimented following high-speed centrifugation. Radioviral ligand was 80% reactive with immobilized Sendai virus antibody, 70% reactive with sheep red blood cells,and 1% reactive with receptor-negative horse red blood cells. Pretreatment of the cells with neuraminidase completely inhibited the virus binding reaction. The reaction with receptor-bearing cells was competitively inhibited by unlabeled Sendai virus, but not by murine leukemia virus or T-2 coliphage. Radioviral ligand binding to human lymphoblastic cell line CCL-119 was a saturable reaction, a result that demonstrates the absence of virus-virus aggregates. The preparation of an effective cell-reactive radioviral ligand was dependent on the initial purification from immobilized receptor-containing proteins. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|